Analysis of Cell Viability in a Live Cell Culture by Fluorescence Microscopy

February 19, 2016

George Washington University, Washington DC
Surgery
Post-doctoral Fellow

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Company:

Thermo Fisher Scientific

Product Name:

ReadyProbes® Cell Viability Imaging Kit, Blue/Green

Catalog Number:

R37609

Cell viability in an ongoing culture is often crucial in determining experimental results obtained from it regarding cell growth, multiplication, turn over rate and apoptosis/necrosis. It becomes important to determine cell viability in a culture as it can affect the overall outcome of any further assays done to observe proliferation, expression or physiological changes as viability certainly affects all these parameters. A good way to check for cell viability in live cultures is by using the assay kit which utilizes a blue and a green dye to differentiate between live and dead cells by fluorescent method.

Experimental Design and Results Summary

Application

Cell culture

Starting Material

Any type of cell that is in culture

Protocol Overview

It is best to do the experiment in a 6 well/12 well plate if there are sufficient cells. Add 2 drops each of NucBlue (stains live cells) and NucGreen (stains cells with compromised ceell membrane/ dead/dying cells) per ml of culture media in each well.keep in the dark (wrap with aluminium foil) and place the plate within a 5% CO2 incubator. Stain for 20-30 mins. depending on your cell type and experimental set-up. It is better to wash off the stained medium after the staining period although it is not mandatory. Observe under fluorescent microscope with DAPI (blue) and FITC (green) filter. Take pictures for a merge of the blue and green channel. The blue cells are the live cells whereas the green are the dead/ dying cells and the ration of blue/ green can be counted.

Tips

Staining time needs optimization

Results Summary

Clear blue or green cells are visible and blue/ green ratio gives the viability of the culture

Additional Notes

None

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Summary

The Good

Easy and reliable method for estimation of cell viability in a live cell culture. Can be utilized to determine cell viability in treatment group or amount of cell death in an inhibitor study experiment.

The Bad

It is recommended not to use the stained cells for any other experimental purpose.

The Bottom Line

Cost effective and reliable ready probes to study cell death/ cell viability in an easy experimental set-up.

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